Elasticity of Phloem Tissues1

Linear displacement transducers and a pressure transducer wereused to simultaneously monitor pressure reduction and shrinkageassociated with phloem exudation following incision of the barkof Fraxinus americana L. The shrinkage associated with the pressurereduction indicated that the sieve tubes had elastic walls.Mathematical models of a mass flow system of phloem transporthave assumed rigid walls for the sieve tubes, but results demonstratedthat the dimensional changes associated with pressure potentialchanges would produce significant differences in volume flowrates if the pressure gradient were maintained at a constantvalue, and thus indicate a requirement for modification of themodels to accommodate elastic cell walls.     

Cell Potentials and Turgor Pressures in Epidermal Cells of Tradescantia and Commelina

Cell membrane potentials have been measured both in epidermalstrips and intact leaf sections of Tradescantia virginiana andCommelina communis, and in epidermal cells over green and overalbino mesophyll cells of T. albiflora var. albovittata. Membranepotentials (_cell) in strips were considerably lower than thosein intact sections and were insensitive to light and to theabsence or presence of calcium. Their response to external cationlevels was indifferent to ionic species. However, in intactleaf sections incubated with calcium present, membrane potentialsresponded to K⁺ levels but not to Na⁺. were more negative thancells in epidermal strips, and responded to changes in illumination. Long-term recordings of _cell and vacuolar K⁺ levels in T. virginianaduring stomatal closure suggest that the fluctuations of _cellwere unrelated to K⁺ movement (which we could not detect) andthus probably to stomatal movement as well. Turgor pressures measured in epidermal cells of intact leafsections of T. virginiana were found to be of the same magnitudeas those previously reported for epidermal strips. It is concludedthat epidermal cells maintain their solute contents during strippingwithout the involvement of an electrophysiological transportsystem. With the possible exception of lateral subsidiary cells,there was no evidence suggesting that ordinary epidermal cellsare capable of osmotic adjustment even when additional KCI wassupplied in the osmoticum. Absolute turgor levels in intactleaf sections kept at constant external KCI were unrelated tosteady state _cell.     

A Comparison of Methods for Measuring Turgor Pressures and Osmotic Pressures of Cells of Red Beet Storage Tissue

Two methods for measuring the turgor pressures of cells in discsof storage tissue of red beet (Beta vulgaris L.) were compared,and a centrifugation method for extracting sap from frozen andthawed tissue was evaluated. Turgor pressures were measureddirectly using a pressure probe, or indirectly using a vapourpressure osmometer. With the latter, discs were placed directlyin the osmometer chamber and turgor was calculated as the differencein osmotic pressure before and after freezing and thawing. Turgorin freshly cut discs, measured with the pressure probe, wasbetween 0-012 MPa and 0.118 MPa with a mean ±s.d. of0.092±;0.032 MPa (n = 24). That measured with the osmometervaried between 0.08 MPa and 0.12 MPa with a mean ±s.d.of 0.09±0.10 MPa (n = 54). After vacuum infiltrationof discs with distilled water, the turgor measured with thepressure probe increased to 1.05–1.12 MPa. Turgor measuredwith the osmometer also increased after vacuum infiltrationbut was, on average, 12% lower than that measured with the pressureprobe. Overall, the results suggest that for routine measurements,the osmometer can provide reasonable estimates of the turgorof cells in beet discs. This is because a number of factorsthat, potentially, could interfere with this method have onlya small effect in this tissue. None of the measured turgorsis indicative of that occurring in intact storage roots becauseboth excision and vacuum infiltration of discs alter the concentrationsof solutes in the extracellular space. The osmotic pressureof sap extracted by centrifugation from frozen and thawed discswas not significantly different from that measured by placingfrozen and thawed discs directly in the osmometer. Solute concentrationsin the sap were not significantly different from those measuredby chemical extraction of discs. Key words: Beta vulgaris, Osmotic pressure, Turgor pressure     

Water Flow Across the Sieve-Tube Boundary: Estimating Turgor and Some Implications for Phloem Loading and Unloading. II. Phloem in the Stem

Confirming a previous analysis by Lang (1974), it is concludedthat in tree trunks, phloem turgor and turgor gradients maybe estimated from osmotic pressure and osmotic-pressure gradients,respectively. The present analysis is an improvement becauseit is based on observed osmotic-pressure gradients rather thansupposed turgor gradients, and allowance is made for sucroseunloading and gradients of external water potential. It is concludedthat the rate of sucrose unloading in tree trunks must be lessthan 50 nmol m^–2 S m^–1. In small plants, higherrates of unloading (100 nmol m m^–2 S m^–1) and steeperconcentration gradients will lead to larger errors, but turgorpressures can still be estimated with acceptable accuracy. Oneshould be more cautious when considering turgor gradients insmall plants, although it seems likely that reasonable estimateswill still be obtained. Assuming plasmodesmatal transport throughan unconstricted cytoplasmic annulus, it is concluded that thesieve elements and their associated cells will sustain verysimilar turgor and osmotic pressures. Convection and diffusioncan both contribute significantly to plasmodesmatal sucroseunloading. Similarly, the plasmodesmatal volume flux will reflecta combination of pressure flow and osmosis. Water fluxes acrossthe sieve element plasmalemma and through the plasmodesmatacan be in opposite directions. It may be possible to assessthe extent of hydraulic coupling between the sieve elementsand their associated cells from studies of phloem water relations Phloem, turgor, osmotic pressure, plasmodesmata, phloem unloading, Munch hypothesis     

Water Flow Across the Sieve-Tube Boundary: Estimating Turgor and Some Implications for Phloem Loading and Unloading. III. Phloem in the Leaf

The theory described in Part I of this series is applied hereto the loading of water and sucrose into the sieve-element-companion-cell(se-cc) complexes of minor veins. It is concluded that symplasticphloem loading cannot account for large increases in osmoticpressure from mesophyll to se-cc complex or the dilution ofsieve tube sap which is evident in leaves. By contrast, loadingfrom the free space into a symplastically isolated se-cc complexcan account for both these observations. Within the se-cc complex,sucrose and water will be transported from the companion cellsto the sieve elements by a pressure-driven flow of solutionthrough the cytoplasmic annuli of plasmodesmata. The associatedchanges in turgor and osmotic pressure are small, and so these-cc complex can be regarded as a single compartment with respectto water potential. The assumption that this compartment isat water flux equilibrium will lead to significant overestimatesof turgor gradients. However, if the trans-membrane water potentialdifference and the external water potential are taken into account,the correlation of such gradients with sieve-element dimensionsand / or transport velocities provides one means of testingthe Munch hypothesis of phloem transport Phloem, turgor, osmotic pressure, plasmodesmata, Münch hypothesis, Phloem loading     

Measurement of Phloem transport rates by an indicator-dilution technique

An indicator-dilution technique for the measurement of flow rates, commonly used by animal physiologists for circulation measurements, was adapted to the measurement of phloem translocation rates in the wheat (Triticum aestivum L.) peduncle. The approach is based on the observation that, during the transport of a given amount of solute, its mean concentration will be inversely proportional to flow rate. For phloem transport in the wheat peduncle, the necessary measurements are (a) the time course of tracer kinetics in the peduncle phloem, (b) the volume of sieve tubes and companion cells in the monitored segment of the peduncle, and (c) the amount of tracer transported past that point. The method was evaluated by in situ monitoring of ³²PO₄ transport in pulse-labeling experiments. Specific activities (i.e.³²P concentrations) of phloem exudate were in good agreement with those calculated from in situ count rates and measured phloem areas. Mass transport rates, calculated from volume flow rates and phloem exudate dry matter content, also agreed well with expected mass transport rates based on measurements of grain growth rate and net CO₂ exchange by the ear. The indicator-dilution technique appears to offer good precision and accuracy for short-term measurements of phloem transport rates in the wheast peduncle and should be useful for other systems as well. In contrast to velocities based on time-delay measurements, it is more precise, more accurate, and produces an estimate of mean, rather than maximum, velocity. Also, since only a single detector is required, it can be applied to very short transport paths.     

Turgor regulation and cytoplasmic free Ca2+ in the algaLamprothamnium

Summary In internodal cells ofLamprothamnium succinctum, turgor regulation in response to hypotonie treatment is inhibited by lowering external Ca²⁺ concentration ([Ca²⁺]_e) from 3.9 (normal) to 0.01 (low) mM. In order to clarify whether a change in the cytoplasmic free Ca²⁺ concentration ([Ca²⁺]_c) is involved in turgor regulation, the Ca²⁺ sensitive protein aequorin was injected into the cytoplasm of internodal cells. A large transient light emission was observed upon hypotonic treatment under normal [Ca²⁺]_e but not under low [Ca²⁺]_e. Thus hypotonic treatment induces a transient increase in [Ca²⁺]_c under normal [Ca²⁺]_e but not under low [Ca²⁺]_e.Abbreviations ASW artificial sea water - _i cellular osmotic pressure - [Ca²⁺]_c cytoplasmic free Ca²⁺ concentration - EDTA ethylenediamine-tetraacetic acid - EGTA ethylenglycol-bis(-aminoethyl ether(N,N-tetraacetic acid - [Ca²⁺]_e external Ca²⁺ concentration - _e external osmotic pressure - GM glass micropipette - GP glass plate - HEPES N-2-hydroxyethylpiperazine-N-2-ethansulfonic acid - MS microscope stage - OL objective lens - PIPES piperazine-N-N-bis(2-ethanesulfonic acid) - W Weight     

High Pressures of Oxygen and Respiration; an Improved Technique and a Demonstration that High Pressures as such may affect the Rate of CO2 Output from plant Tissues1

A technique is described which allows the metabolic rate ofCO₂ output to be determined during the first 24 hours of treatmentwith oxygen at high pressures. With potatoes, and to a lesser extent with apples, the rateof CO₂ output was inhibited by an increase of the absolute pressureto 3 atmospheres, the partial pressure of oxygen remaining at20 per cent, of 1 atmosphere. The inhibition is attributed tomechanical distortion of the tissue caused by the increase inpressure.     

An Instrument for Non-Destructive Measurement of the Pressure Potential (Turgor) of Leaf Cells

An instrument is described which permits the non-destructivemeasurement of the mean pressure potential (turgor)of leaf laminacells. Calibration shows that the instrument gives a voltageoutput which is linearly related to mean pressure potentialof living leaf cells as determined with a pressure chamber.Measurements may be made very quickly in the field or controlledenvironment with a resolution of at least 50 k Pa.     

Response of Tissues2

Efflux of K⁺ from tomato fruit pericarp disks was not affectedby 100 µM tomatine, and only slightly by 1 mM. No differentialresponses to the alkaloid were observed in disks obtained fromfruits at different stages of development, and there was noevidence of any correlation between the level of endogenoustomatine and the effect of exogenous tomatine. Growth of tomatoseedlings in liquid nutrient medium was unaffected by 1 mM tomatinewhereas the growth oflettuce seedlings was stunted in the presenceof 100 µM alkaloid. The polyene antibiotic nystatin gavesimilar resultsto tomatine, having little effect on tomato tissuesbut causing marked inhibition of lettuce seedling development.     

改良的半叶法测定茶光合速率时韧皮部处理方法的比较

采用环割、ＴＣＡ（三氯醋酸）化学抑制、石蜡烫伤茶叶叶柄韧皮部,以阻止光合产物外运,并通过石蜡切片观察韧皮部破坏程度与其光合速率的关系。结果表明：环割能有效去除韧皮部,阻止光合产物运输,光合速率高;其次TCA 抹,能致使输导系统筛管塞堵子,光合产穆能外运;最后是石蜡烫伤,产生结晶簇结输导组织和运输带来了障碍。     

Solution-Flow in the Phloem: II. Phloem Transport of THO in Beta vulgaris

Translocation profiles along the path were studied using a modified flap-feeding technique for the simultaneous application of THO and ¹⁴C-sucrose. A re-evaluation of a mathematical model for phloem transport with reversible lateral exchange of tracer along the path indicates that lower apparent velocities for THO as compared to labeled carbohydrate are primarily due to extensive lateral exchange of THO along the conduction path. Path-chilling experiments support the concept that THO and ¹⁴C-sucrose exhibit different lateral exchange characteristics. The data presented are consistent with a solutionflow mechanism.     

Phloem Glutamine and the Regulation of O2 Diffusion in Legume Nodules

The aim of the present study was to test the hypothesis that the N content or the composition of the phloem sap that supplies nodulated roots may play a role in the feedback regulation of nitrogenase activity by increasing nodule resistance to O2 diffusion. Treating shoots of lupin (Lupinus albus cv Manitoba) or soybean (Glycine max L. Merr. cv Maple Arrow) with 100 [mu]L L-1 NH3 caused a 1.3-fold (lupin) and 2.6-fold (soybean) increase in the total N content of phloem sap without altering its C content. The increase in phloem N was due primarily to a 4.8-fold (lupin) and 10.5-fold (soybean) increase in the concentration of glutamine N. In addition, there was a decline in both the apparent nitrogenase activity and total nitrogenase activity that began within 4 h and reached about 54% of its initial activity within 6 h of the start of the NH3 treatment. However, the potential nitrogenase activity values in the treated plants were not significantly different from those of the control plants. These results provide evidence that changes in the N composition of the phloem sap, particularly the glutamine content, may increase nodule resistance to O2 diffusion and, thereby, down-regulate nodule metabolism and nitrogenase activity by controlling the supply of O2 to the bacteria-infected cells.     

Measurement of the Water Potential of Intact Plant Tissues1

A modified design of a thermocouple psychrometer is describedwhich utilizes the Peltier effect for producing an exceedinglysmall wet-bulb thermometer. The thermocouple assembly has beenreduced to miniature dimensions and the thermocouple chamberconsists of a silver cylinder (volume 0.06 cm³) whose base isformed by the material under observation. This makes it possibleto measure water potentials over limited surfaces of intactplant tissues, e.g. germinating pea seeds (Pisum sativum L.).The apparatus is capable of measuring water potentials downto a magnitude of about –6,000 joules/kg.³